2016-11-19T15:57:02ZDual Quorum Quenching Capsules: Disrupting two bacterial communication pathways that lead to virulencehttp://hdl.handle.net/1903/18745
Dual Quorum Quenching Capsules: Disrupting two bacterial communication pathways that lead to virulence
Rhoads, Melissa Katherine
Healthcare Associated Infections (HAIs) in the United States, are estimated to cost nearly $10 billion annually. And, while device-related infections have decreased, the 60% attributed to pneumonia, gastrointestinal pathogens and surgical site infections (SSIs) remain prevalent. Furthermore, these are often complicated by antibacterial resistance that ultimately cause 2 million illnesses and 23,000 deaths in the US annually.
Antibacterial resistance is an issue increasing in severity as existing antibiotics are losing effectiveness, and fewer new antibiotics are being developed. As a result, new methods of combating bacterial virulence are required. Modulating communications of bacteria can alter phenotype, such as biofilm formation and toxin production. Disrupting these communications provides a means of controlling virulence without directly interacting with the bacteria of interest, a strategy contrary to traditional antibiotics.
Inter- and intra-species bacterial communication is commonly called quorum sensing because the communication molecules have been linked to phenotypic changes based on bacterial population dynamics. By disrupting the communication, a method called ‘quorum quenching’, bacterial phenotype can be altered. Virulence of bacteria is both population and species dependent; each species will secrete different toxic molecules, and total population will affect bacterial phenotype9. Here, the kinase LsrK and lactonase SsoPox were combined to simultaneously disrupt two different communication pathways with direct ties to virulence leading to SSIs, gastrointestinal infection and pneumonia.
To deliver these enzymes for site-specific virulence prevention, two naturally occurring polymers were used, chitosan and alginate. Chitosan, from crustacean shells, and alginate, from seaweed, are frequently studied due to their biocompatibility, availability, self-assembly and biodegrading properties and have already been verified in vivo for wound-dressing. In this work, a novel functionalized capsule of quorum quenching enzymes and biocompatible polymers was constructed and demonstrated to have dual-quenching capability. This combination of immobilized enzymes has the potential for preventing biofilm formation and reducing bacterial toxicity in a wide variety of medical and non-medical applications.
2016-01-01T00:00:00ZEffects of Load History on Ovine Spinal Range of Motionhttp://hdl.handle.net/1903/18711
Effects of Load History on Ovine Spinal Range of Motion
Rotunno, Giuliana
Loading of spinal motion segment units alters biomechanical properties by modifying flexibility and range of motion. This study utilizes angular displacement due to an applied bending moment to assess biomechanical function during high-magnitude and prolonged compressive loading of ovine lumbar motion segments. High compressive loads, representative of physiological lifestyle and occupational behaviors, appear to limit fluid recovery of the intervertebral disc, thereby modifying spinal flexibility and increasing spinal instability. Intermittent extensions, or backwards bending movements, may provide a protective effect against the load-induced spinal instability. This study contributes a greater understanding of the effects of load history on the function and health of the lumbar spine. Findings may inform future efforts investigating adjustments in spinal posture to preserve or promote the recovery of lumbar spinal biomechanics.
2016-01-01T00:00:00ZDesign and Pilot Study for an Efficient High-Throughput Automated Computer-Vision Guided Intelligent De-Calyxing Machine for Post-Harvest Strawberry Processinghttp://hdl.handle.net/1903/18695
Design and Pilot Study for an Efficient High-Throughput Automated Computer-Vision Guided Intelligent De-Calyxing Machine for Post-Harvest Strawberry Processing
Lin, John
Strawberries harvested for processing as frozen fruits are currently de-calyxed manually in the field. This process requires the removal of the stem cap with green leaves (i.e. the calyx) and incurs many disadvantages when performed by hand. Not only does it necessitate the need to maintain cutting tool sanitation, but it also increases labor time and exposure of the de-capped strawberries before in-plant processing. This leads to labor inefficiency and decreased harvest yield. By moving the calyx removal process from the fields to the processing plants, this new practice would reduce field labor and improve management and logistics, while increasing annual yield.
As labor prices continue to increase, the strawberry industry has shown great interest in the development and implementation of an automated calyx removal system. In response, this dissertation describes the design, operation, and performance of a full-scale automatic vision-guided intelligent de-calyxing (AVID) prototype machine. The AVID machine utilizes commercially available equipment to produce a relatively low cost automated de-calyxing system that can be retrofitted into existing food processing facilities.
This dissertation is broken up into five sections. The first two sections include a machine overview and a 12-week processing plant pilot study. Results of the pilot study indicate the AVID machine is able to de-calyx grade-1-with-cap conical strawberries at roughly 66 percent output weight yield at a throughput of 10,000 pounds per hour.
The remaining three sections describe in detail the three main components of the machine: a strawberry loading and orientation conveyor, a machine vision system for calyx identification, and a synchronized multi-waterjet knife calyx removal system. In short, the loading system utilizes rotational energy to orient conical strawberries. The machine vision system determines cut locations through RGB real-time feature extraction. The high-speed multi-waterjet knife system uses direct drive actuation to locate 30,000 psi cutting streams to precise coordinates for calyx removal.
Based on the observations and studies performed within this dissertation, the AVID machine is seen to be a viable option for automated high-throughput strawberry calyx removal. A summary of future tasks and further improvements is discussed at the end.
2016-01-01T00:00:00ZMICROFLUIDIC ASSAY PERFORMANCE ENHANCEMENT USING POROUS VOLUMETRIC DETECTION ELEMENTS FOR IMPEDEMETRIC AND OPTICAL SENSINGhttp://hdl.handle.net/1903/18662
MICROFLUIDIC ASSAY PERFORMANCE ENHANCEMENT USING POROUS VOLUMETRIC DETECTION ELEMENTS FOR IMPEDEMETRIC AND OPTICAL SENSING
Wiederoder, Michael
Microfluidic technologies have great potential to help create automated, cost-effective, portable devices for rapid point of care (POC) diagnostics in diverse patient settings. Unfortunately commercialization is currently constrained by the materials, reagents, and instrumentation required and detection element performance. While most microfluidic studies utilize planar detection elements, this dissertation demonstrates the utility of porous volumetric detection elements to improve detection sensitivity and reduce assay times.
Impedemetric immunoassays were performed utilizing silver enhanced gold nanoparticle immunoconjugates (AuIgGs) and porous polymer monolith or silica bead bed detection elements within a thermoplastic microchannel. For a direct assay with 10 µm spaced electrodes the detection limit was 0.13 fM AuIgG with a 3 log dynamic range. The same assay was performed with electrode spacing of 15, 40, and 100 µm with no significant difference between configurations. For a sandwich assay the detection limit was10 ng/mL with a 4 log dynamic range. While most impedemetric assays rely on expensive high resolution electrodes to enhance planar senor performance, this study demonstrates the employment of porous volumetric detection elements to achieve similar performance using lower resolution electrodes and shorter incubation times.
Optical immunoassays were performed using porous volumetric capture elements perfused with refractive index matching solutions to limit light scattering and enhance signal. First, fluorescence signal enhancement was demonstrated with a porous polymer monolith within a silica capillary. Next, transmission enhancement of a direct assay was demonstrated by infusing aqueous sucrose solutions through silica bead beds with captured silver enhanced AuIgGs yielding a detection limit of 0.1 ng/mL and a 5 log dynamic range. Finally, ex situ functionalized porous silica monolith segments were integrated into thermoplastic channels for a reflectance based sandwich assay yielding a detection limit of 1 ng/mL and a 5 log dynamic range. The simple techniques for optical signal enhancement and ex situ element integration enable development of sensitive, multiplexed microfluidic sensors.
Collectively the demonstrated experiments validate the use of porous volumetric detection elements to enhance impedemetric and optical microfluidic assays. The techniques rely on commercial reagents, materials compatible with manufacturing, and measurement instrumentation adaptable to POC diagnostics.
2016-01-01T00:00:00Z